title: A novel hepatitis C virus NS5B polymerase assay of de novo initiated RNA synthesis directed from a heteropolymeric RNA template.
abstract: The hepatitis C virus (HCV) NS5B RNA-dependent RNA polymerase is essential for viral replication and a clinically validated antiviral target. Discovery of HCV polymerase inhibitors is greatly facilitated by the availability of a suitable biochemical assay using purified NS5B. We describe here a novel NS5B polymerase assay of de novo initiated RNA synthesis directed from a synthetic heteropolymeric RNA template modified with dideoxycytidine at its 3'-end. This assay has been successfully used for screening and characterization of both initiation and elongation inhibitors of the HCV NS5B polymerase.
authors.affiliation: Department of Infectious Diseases, Merck Research Laboratories, Kenilworth, NJ, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Ferrari
authors.forename: Hsueh-Cheng
authors.initials: HC
authors.lastname: Huang
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D057075:Enzyme Assays / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016174:Hepacivirus / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D012367:RNA, Viral / Q000096:biosynthesis*; D013698:Templates, Genetic; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014779:Virus Replication; D012324:RNA-Dependent RNA Polymerase
chemical_list: D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase